Assessment of Thermophilic Campylobacter Load in Surface Water Bodies of Uttarakhand Province of India Through Real-time Quantitative Polymerase Chain Reaction (qpcr) Assay

Background: Being the leading cause of human diarrhoea globally, Campylobacter gained importance in developed and developing countries in the last few decades. It causes a foodborne illness that generally spreads through eating unprocessed foods of animal origin, contact with untreated water and animal feces. Contaminated water with Campylobacter can be a potential source of its spread into the human population. Therefore, it is of utmost importance to estimate the Campylobacter load of surface water bodies like rivers and lakes. The advancement in molecular techniques like quantitative real-time PCR (qPCR) has made it quite convenient to quantify the pathogen load by amplification of its genomic DNA isolated directly from an environmental sample. The present study aimed to assess the Campylobacter load in surface water bodies of the Uttarakhand Province of India using a qPCR assay. Method: A total of 160 water samples were collected from different surface water bodies located in the human vicinity, including lakes, rivers, dams, industrial drains, irrigated fields and recreational waters from the Uttarakhand province of India. The Campylobacter spp. was isolated from the samples and characterized using biochemical methods and molecular confirmation using PCR. Further, the bacterial load of water samples collected from different sources was determined by SYBR dye-based Real-Time qPCR assay using absolute quantitation following the standard curve method. Result: The biochemical and molecular investigations confirm the presence of Campylobacter spp. in the water samples. The bacterial load was found between a minimum of 7.7 x 102 cells to a maximum of 2.7 x 105 cells in water samples. The higher bacterial load in stagnant water sources near areas of human-animal interface suggests contamination of water bodies with human and animal fecal waste, posing a significant public health concern.